PCR技术检测海运船舶压载水病原弧菌初探

船舶压载水引起的外来物种入侵,及由此引发的经济、环境和卫生等方面的灾害性后果,已成为当前世界海洋所面临的最大威胁之一.弧菌作为一种广泛存在于海洋环境的病原微生物,其数量已成为船舶压载水是否可以排放的一项指标.本文采用PCR扩增病原弧菌特征基因技术,结合弧菌相关16S rRNA基因文库,初步探讨压载水中病原弧菌快速检测技术.从采集自舟山远洋船舶压载水水样S1中,检测到hlyA和vvh基因,表明样品中含有霍乱弧菌(Vibrio cholerae)和创伤弧菌(V.vulnificus)的毒力基因.文库序列分析结果显示,重组克隆子中部分序列与副溶血弧菌(V.parahaemolyticus)和费氏弧菌(V.fischeri)相似性较高.研究结果表明,压载水样品S1中含有多种对人和动植物有害的病原弧菌,对海洋生态平衡、公共卫生安全和海洋养殖业具有潜在危害.

Detection of pathogenic Vibrio in the ballast water by polymerase chain reaction

The potential of ballast water to act as a major vector for invasive alien species has long been recognised.Invasive alien species caused by ballast water frequently outcompete or prey on native species and spread diseases to plants,animals,and human.They often encroach on,damage,or degrade assets and result in significant economic impacts.Vibrios are highly abundant in marine environments.Some of them such as Vibrio cholerae,V.vulnificus and V.parahaemolyticus are pathogenic for human being.An international ballast water performance standard,which prescribed Vibrio cholerae（O1 and O139） with less than 1cfu per 100 mL,was adopted by the International Maritime Organization（IMO） in 2004.Multiple pathogens in ballast water samples from Zhoushan archipelago located in the eastern of China were detected.Two virulence-associated genes（hlyA and vvh） usually from V.cholerae and V.vulnificus were detected by polymerase chain reaction.The molecular library containing 16S rRNA gene of Vibrio was constructed subsequently.The results indicated that some V.parahaemolyticus-and V.fischeri-like strains existed in the ballast water sample S1.It was concluded that the ballast water sample contained some pathogenic Vibrios.This paper presented new molecular data based on virulence-associated gene and LSU-rDNA sequences that demonstrate a rapid and effective method for detecting pathogenic Vibrio in the ballast water.